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Technologies for tissue culture electrophysiologic recording

Author

Lorne Bonkowski, University of the Pacific

Date of Award

1973

Document Type

Thesis

Degree Name

Master of Science (M.S.)

Department

Pharmacy

First Advisor

Howell I. Runion

First Committee Member

R. E. H[?]

Second Committee Member

Donald M. Pace

Third Committee Member

Donald Y. Shirachi

Abstract

The development of tissue culture electrophysiology has been hindered by these numerous technological problems. The function of this thesis was to identify and propose solutions to these technological problems and concurrently develop a system for prolonged intracellular GME monitoring (30 to 40 hours) of membrane parameters from cells grown in tissue culture.

The technological problems identified in the present study and for which solutions have been proposed are: (1) Maintenance of a sterile environment for an open tissue culture vessel in which GME measurements can be made for periods exceeding 30 to 40 hours.; (2) Elimination of environmental electrical interference which normally interferes with GME recording and data displaying.; (3) Design of an optical instrumentation system that is compatible with microelectrode penetration needs and tissue culture cell observation.; (4) Elimination of building and peripheral instrument vibrational influences at the microelectrode recording site.; (5) A method of microelectrode preparation that would produce consistent glass microelectrodes meeting the peculiar needs of tissue culture electrophysiology.; (6) Design of a culture recording chamber which would permit: (a) culturing of cells using standard tissue culture techniques.; (b) easy access of cells for microelectrode penetration.; (c) continuous optical observation.; (d) continuous temperature regulation of cultured cells for periods exceeding 30 to 40 hours.; (e) shielding against electrical field disturbances during microelectrode recording.; (f) rapid perfusion of media to the chamber without causing harsh turbulence allowing continuous intracellular recording via glass microelectrode even during perfusion of substances.; (g) microelectrodes penetration of cells over a wide range of angles.; (7) Design of a complete intracellular tissue culture electrophysiology monitoring system.; and (8) Development of explant and culturing procedures to provide cell substrate to test the effectiveness of the tissue culture electrophysiology system design.

Pages

57

Recommended Citation

Bonkowski, Lorne. (1973). Technologies for tissue culture electrophysiologic recording. University of the Pacific, Thesis. https://scholarlycommons.pacific.edu/uop_etds/1828